During the next year we will focus studies of globin gene regulation in three areas: 1) 5'-capping and methylation of globin precursor RNA; 2) normal transcription rates of alpha and beta globin genes; and 3) globin gene structure in the alpha-thalassemia syndromes in man. First, we propose to characterize 5'-capping and both 5'-end methylation and internal methylation of globin precursor RNA in murine erythroid precursor cells labeled with (3H-methyl) methionine. 5'-end methylation of mature nuclear globin RNA will be compared to that of cytoplasmic globin RNA. Second, rates of alpha and beta globin RNA transcription will be compared in murine erythroblasts to determine whether regulation of the alpha/beta ratio occurs in transcription. Both initiation and elongation of alpha and beta transcripts will be studied. Third, using restriction-endonuclease-digested DNA and molecular hybridization techniques, we will study alpha globin gene structure in Black and Oriental individuals with alpha-thalassemia. In summary, we will elucidate further how normal and mutant globin genes are regulated at the levels of gene number, transcription, and posttranscriptional processing.